• zebrafish system

Anti-Grouper PAb IgM-H (Hot Product)

Product Name:

Rabbit polyclone antibody against grouper(E. coioides) IgM H chain Puriified Ig, lyophiliczed protein

Catalog Number: GDN‐A0004

Size: 50ug, lyophilised protein

Reconstitution: Add 200 μl of distilled water

Lot Number: See label on vial

Host: Rabbit polyclonal

Species reactivity: Grouper (E.Coioides)

Cross‐reactive species:

  • Epinephelus coioides × Epinephelus malabaricus (青斑)
  • Epinephelus coioides × Epinephelus lanceolatus (青龍班)
  • Epinephelus lanceolatus (龍膽石斑)
  • Epinephelus lanceolatus × Epinephelus fuscoguttatus (龍虎斑)
  • Epinephelus tukula (藍身大石斑)
  • Cromileptes altivelis (駝背鱸、老鼠斑)

Application:

  • Western Blot : 20000X
  • ELISA: 20000X
  • Immunocytochemistry (ICC): 10000X

ELISA:

  1. Coating: Coat 5μg of purified gIgM protein in the ELISA plate at 37℃ for 2 hours.
  2. Blocking: Block with gelatin‐NET solution at room temperature for 1~2 hours. Wash with PBST for three times.
  3. First antibody: Incubate with rabbit anti‐gIgM serum in adequate concentration (dilute with gelatin‐NET solution) at 37℃ for 30 min and 4℃ for 1 hour. Wash with PBST for three times.
    (Pre‐immune serum: 2000X, immune serum: 2000, 4000, 8000X)
  4. Secondary antibody: Incubate with goat anti‐rabbit Ig conjugated with HRP inadequate concentration (2000X) at 37℃ for 30 min and 4℃ for 1 hour. Wash with PBST for five times.
  5. Substrate solution: Incubate with OPD substrate solution at 37℃ for 30 min. Measure the absorbance at 405 nm wavelengths and 650 nm wavelengths as background by ELISA reader.

Ig against grouper IgM H chain (gIgM H) was purified form the rabbit antiserum and lyophilized. Therefore, we recommend to dissolve the lyophilized Ig in 200μL ddH2O and dilute it for about 20,000~40,000 fold.

 

Description:
The optimal dilution factor of rabbit antiserum against grouper IgM H‐chain in western blot was no less than 8000‐40000X. In a separate assay, the optimal dilution factor of antiserum for ELISA was 20000X (data not shown).

Application Notes:
The following concentration ranges are recommended starting points for this product. The investigator should determine the optimal working concentrations for specific applications.

Storage:
For longer periods of storage, store at ‐20°C. Avoid repeat freeze‐thaw cycles.

This product is for in vitro research use only.

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